The control or SENP1-aP2KO mice were fed with normal chow, water consumption and food intake was recorded daily. For the glucose tolerance test (GTT) and insulin tolerance test (ITT), the mice were fasted overnight, followed by an intraperitoneal injection of glucose (GTT; 1 g/kg body weight) or insulin (ITT; 0.75 U/kg body weight). Blood glucose levels at various times after the injections were determined by an electronic glucometer (Abbott Diabetes Care, Alameda, CA). Lipids analysis and lipoprotein profile measurement were performed 60 (link). Briefly, mice were fasted for 12–14 hrs before blood samples were collected by retro-orbital venous plexus puncture. Plasma was separated by centrifugation and stored at −80°C. Total plasma cholesterol, triglycerides and free fatty acids (FFA) were enzymatically measured with the Amplex red cholesterol assay kit (Molecular Probes), serum triglyceride determination kit and free fatty acid kit (Sigma), respectively, according to the manufacturer’s instructions. The lipid distribution in plasma lipoprotein fractions were assessed by fast-performance liquid chromatography (FPLC) gel filtration with two Superose 6 HR 10/30 columns (Pharmacia).
Electronic glucometer
Manufactured by Abbott
Sourced in United Kingdom, United States
The electronic glucometer is a compact, portable device designed to measure and display the user's blood glucose level. It utilizes a small test strip inserted into the device, which interacts with a drop of blood to determine the glucose concentration. The glucometer provides a digital readout of the measurement, allowing users to monitor their blood sugar levels accurately and conveniently.
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4 protocols using electronic glucometer
1
Metabolic Profiling of SENP1-aP2KO Mice
2
Endothelial SENP1 Knockout Diabetic Mouse Model
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Vascular endothelial cell (EC)-specific deletion of SENP1 (SENP1-ECKO), which was generated by mating SENP1lox/lox with VE-cadherin-Cre mice24 (link),25 (link) followed by breeding to C57BL/6 background. The deletion of SENP1 in SENP1-ECKO was verified by qRT-PCR using primers amplifying exons 5–6 and specific Cre. SENP1lox/lox mice were used as controls. We used a standard protocol of STZ injections to induce type I diabetes (T1DM) in C57BL/6 mice. Briefly, C57BL/6 mice at 8 weeks of age mice were injected intraperitoneally with 45 mg/kg STZ or PBS daily for 4 days. Blood glucose concentrations in non-fasting mice were measured 14–16 days after the first injection by an electronic glucometer (Abbott Diabetes Care, Alameda, CA). Blood glucose levels in STZ-treated mice were 350 ± 40 mg/dL (higher than 250 mg/dL were considered diabetic). PBS-treated mice were used as controls for further experiments. Mice were cared for in accordance with National Institutes of Health guidelines, and all procedures were approved by the Yale University Animal Care and Use Committee.
3
Neonatal Calf Blood Metabolite Profiling
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Calves´ blood samples were taken from jugular vein at birth, 55, 98, 153, and 236 days. Samples were taken at the same time in the day after 16 h fasting and no water supply, preserved in ice for 3 h, centrifuged (2500× g, 15 min) for serum collection and stored at −20 °C until laboratory analysis. Glucose level was measured using an electronic glucometer (Abbott©, Berkshire, UK) immediately after the blood sample was taken [17 (link)]. Insulin concentration was assessed in serum samples by radioimmunoassay technique (RIA) using anti-bovine insulin antibody (Sigma, St. Louis, MO, USA) and standard human insulin (Laboratorios Beta, Buenos Aires, Argentina); the minimum detectable concentration was 0.05 ng/mL. Intra- and inter-assay coefficients of variation were <7.8% and 10.3%, respectively. Serum IGF1 concentration was serum quantified in one assay via RIA after acid ethanol extraction [18 (link)] using IGF1 antibody (UB2-495) of the NIDDK. Intra-assay coefficient of variation was 7.6%, and the minimum detectable concentration was 2.4 ng/mL.
4
Metabolic Profiling in SENP1-Deficient Mice
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The control or SENP1-aP2KO mice were fed with normal chow, water consumption and food intake was recorded daily. For the GTT and ITT, the mice were fasted overnight, followed by an intraperitoneal injection of glucose (GTT; 1 g kg−1 body weight) or insulin (ITT; 0.75 U kg−1 body weight). Blood glucose levels at various times after the injections were determined by an electronic glucometer (Abbott Diabetes Care, Alameda, CA, USA). Lipids analysis and lipoprotein profile measurement were performed60 (link). In brief, mice were fasted for 12–14 h before blood samples were collected by retro-orbital venous plexus puncture. Plasma was separated by centrifugation and stored at −80 °C. Total plasma cholesterol, TGs and FFAs were enzymatically measured with the Amplex red cholesterol assay kit (Molecular Probes), serum triglyceride determination kit and free fatty acid kit (Sigma), respectively, according to the manufacturer's instructions. The lipid distribution in plasma lipoprotein fractions were assessed by fast-performance liquid chromatography gel filtration with two Superose 6 HR 10/30 columns (Pharmacia).
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